李晓华,黄粤,马婷婷,皮婷,姚家成,夏珍珍,冯喆,温馨.具有抗稻瘟病菌活性的链霉菌ⅠT2菌株的分离鉴定及其attB位点的分析[J].中南民族大学学报自然科学版,2019,(2):193-198
具有抗稻瘟病菌活性的链霉菌ⅠT2菌株的分离鉴定及其attB位点的分析
Isolating and identifying of Streptomyces sp.ⅠT2 with antifungal activity to Magnaporthe Grisea and analysis of attB site
  
DOI:10.12130/znmdzk.20190208
中文关键词: 稻瘟病菌  链霉菌ⅠT2菌株  attB位点  位点特异性重组
英文关键词: Magnaporthe grisea  Streptomyces sp. IT2 strain  attB site  site specific recombination
基金项目:国家自然科学基金资助项目(31070087;湖北省自然科学基金重点项目(2011CDA079);中央高校基本科研业务费专项(CZY19018)
作者单位
李晓华,黄粤,马婷婷,皮婷,姚家成,夏珍珍,冯喆,温馨 中南民族大学 生命科学学院 微生物资源与利用湖北省工程技术研究中心武汉 430074 
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中文摘要:
      以稻瘟病菌NO-1为指示菌,从神农架地区土壤中筛选得到一株微生物,命名为ⅠT2. 通过形态、生理生化研究和16S rDNA基因序列同源性比对,初步鉴定为链霉菌属. 通过不同发酵条件对链霉菌ⅠT2菌株发酵液抑菌活性的影响研究,链霉菌ⅠT2菌株较适合的发酵条件为:发酵温度为26.5~30 ℃,发酵转速为150 r/min,发酵时间为48 h. 利用PCR扩增获得ⅠT2菌株的attB位点序列,链霉菌ⅠT2菌株的attB位点序列长277 bp,它与S. griseus M095 的attB 位点核心区域的序列的相似度最高,达到97%,具有发生交换的核心区域5'-TT. 含有ΦC31attP 位点的整合型质粒pSET152可通过attP位点与链霉菌ⅠT2菌株的染色体上的attB位点发生位点特异性重组.
英文摘要:
      A strain of microorganism was screened from the soil of Shennongjia area with Magnaporthe grisea NO-1 as indicator, which was named as IT2. It was identified as Streptomyces by morphological, physiological, biochemical studies and 16S rDNA gene sequence. The effect of different culture conditions on the antibacterial activity of the IT2 strain fermentation broth was determined by cup and saucer method. The results showed that the fermentation conditions of Streptomyces sp. IT2 strain were as follows: the fermentation temperature was 26.5-30 °C, the fermentation speed was 150 r/min, and the fermentation time was 48 h. The attB locus sequence of the IT2 strain was obtained by PCR amplification. The attB locus sequence of the Streptomyces sp. IT2 strain was 277 bp in length, which had the highest similarity to the sequence of the attB locus core region of S. griseus M095, reaching 97%. The integrative plasmid pSET152 including ΦC31 attP site recombined specifically between the attP site in pSET152 and the attB site in the chromosome of Streptomyces sp. IT2 strain.
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